CN114874234A - Tricyclic compound serving as KRAS G12C inhibitor and application thereof - Google Patents

Abstract

The invention provides a novel tricyclic compound shown in a formula (I) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the tricyclic compound and an application of the tricyclic compound or the pharmaceutically acceptable salt thereof in preparation of medicines for preventing and/or treating KRAS G12C mutant tumors.

Description

Tricyclic compound serving as KRAS G12C inhibitor and application thereof

The invention claims priority of prior application with patent application number 202110162783.3 entitled "tricyclic compound as KRAS G12C inhibitor and its application" filed by the intellectual Property office of China at No. 02/05/2021. The entire contents of the above-mentioned prior application are incorporated herein by reference.

Technical Field

The invention relates to a novel tricyclic compound or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the novel tricyclic compound or the pharmaceutically acceptable salt thereof and application of the novel tricyclic compound or the pharmaceutically acceptable salt thereof as a KRAS G12C inhibitor in preventing or treating related diseases.

Background

Ras is the first human protooncogene to be found and has three family members, Hras, Kras and Nras (Barbacid M, Annu Rev Biochem, 1987; 56: 779-827). RAS, a small G protein, has GTP hydrolase activity, is localized to the inner side of the cytoplasmic membrane, and its activity is regulated by differential binding to GTP/GDP. It is activated (on) when it binds to GTP and is in the inactive state (off) when it binds to GDP. RasGTP kinases are crucial in many signal networks, and have the function of signal integration and transmit signals to downstream effectors, and participate in vital activities such as cell movement, cytoskeleton assembly, vesicle and nuclear transport, and further regulate vital processes such as cell proliferation, differentiation, senescence and apoptosis (Fernandez-Medarde A, et al, Genes Cancer, 2011; 2(3): 344-58). Therefore, RAS proteins are considered as important molecular switch proteins in cell signaling.

The protooncogene Ras becomes an oncogene having oncogenic activity after being activated. Ras gene activation modes include point mutation, overexpression and insertion activation. Among them, the most common way in which Ras genes are activated is by point mutation, and carcinogenesis is mainly achieved by means of point mutation. It has now been found that 151 different Ras point mutations, mainly focused on glycine at positions 12, 13 and glutamine at position 61 (Prior IA, et al, Cancer Res 2012; 72(10): 2457-67). Among the above mutations, the G12 point mutation is most common, and the G12 mutation is predominant in KRAS and Hras. Among KRAS mutations, it has been found that there are 15 different point mutations for G12, including G12A, G12D, G12F, G12K, G12N, G12S, G12V, G12Y, G12C, G12E, G12I, G12L, G12R, G12T, and G12W. Among them, the G12D mutation accounted for about 41%, 28% and 14% of the total G12 mutations, G12V and G12C (Hobbs GA, et al, Cancer Cell, 2016; 29(3): 251-3).

Ras mutations are an important cause of promotion of the development of a variety of cancers, often occurring in the early stages of tumorigenesis. These mutation-activated RAS proteins are subject to uncontrolled cell growth and proliferation. Statistically, the 5 cancers with the highest Ras mutation frequency are ductal pancreatic adenocarcinoma, colorectal Cancer, multiple myeloma, lung Cancer and skin melanoma, and the mutation frequencies are 97.7%, 52.2%, 42.6%, 32.2% and 29.1%, respectively (Albertii AF, et al, Bull Cancer, 2017; 104(7/8): 662-74). Notably, in these Ras mutations, the frequency of KRAS mutations was significantly higher than the other two mutations. For example, in pancreatic ductal adenocarcinoma, KRAS mutation rates were as high as 97.7%, while Nras and Hras were all 0; in colorectal cancer, the KRAS mutation rate was as high as 44.7% in 52.2% of Ras mutation rates. In studies on non-small cell lung cancer, Ras mutations were found to occur mostly at codon 12, most commonly at the G12C point, followed by G12V and G12D (Yoon YK, et al, Mol Carcinog 2010; 49(4): 353-62).

The activating mutation of KRAS gene is closely related to the generation and development of human malignant tumor and the recurrence of tumor. Genetic and biochemical studies have demonstrated that KRAS-dependent signaling plays an important role in regulating the growth, proliferation, invasion and metastasis of a variety of cancer cells. KRAS is also considered a marker in patient prognosis. KRAS-mutated patients have a shorter survival in patients with non-small cell lung cancer than KRAS wild-type patients, particularly patients with a G12C point mutation (Svaton M, et al, Anticancer Res, 2016; 36(3): 1077-82). Therefore, the scientific community is always dedicated to searching for small molecules capable of combining with specific mutant KRAS proteins and inhibiting the activation and function of KRAS proteins, thereby preventing the transduction of downstream signaling pathways thereof and finally playing a role in inhibiting the growth of tumors.

KRAS G12C small molecule inhibitors were first reported in the Nature journal by the Shokat topic group of Howde Houston medical institute in 2013 (Ostrem J M, et al, Nature,2013,503(7477): 548-. These inhibitors can bind to the allosteric binding pocket below the region of switch ii of the KRAS protein molecule and form a covalent bond with nearby Cys12, thereby selectively inhibiting activation of KRAS G12C. Based on the above studies, a new KRAS G12C irreversible covalent inhibitor ARS-853 was reported by Wellspring, 2016, to the KRAS G12C molecular switch II allosteric pocket, which can immobilize KRAS protein in an inactive GDP-binding conformation and have selective inhibitory activity against a variety of KRAS G12C mutation-positive tumor cells (Patricelli M P, et al, Cancer Discov,2016,6(3): 316-.

At present, attempts have been made to identify heterocyclic compounds as covalent inhibitors of KRAS G12C including benzoxazinones, pyridopyrimidinones and tetrahydropyridopyrimidines and the like (WO2017/201161, WO2018/119183, WO2018/217651, WO2018/206539, WO2018/143315, WO2017/087528, WO 2020/282 081, WO/2020/178282). The above compounds are all described as KRAS inhibitors for the treatment of tumors. In view of the large tumor market and unmet market demand, the development of KRAS G12C inhibitors has a great market prospect.

Disclosure of Invention

The invention provides a compound shown as a formula (I) or a pharmaceutically acceptable salt thereof:

wherein,

R ¹ selected from the group consisting of ^1a Substituted of the following groups: c ₆ -C ₁₀ Aryl, 5-10 membered heteroaryl;

R ² 、R ³ independently selected from H, F, Cl, Br, I, CN, OH or optionally substituted with R ^2a Substituted of the following groups: NH (NH) ₂ 、C ₁ -C ₆ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7 membered heterocyclyl, C ₁ -C ₆ Alkoxy radical, C ₃ -C ₆ Cycloalkyloxy, 4-7 membered heterocyclyloxy, C ₂ -C ₆ Alkenyl radical, C ₂ -C ₆ An alkynyl group;

R ^4a 、R ^4b selected from H, or R ^4a 、R ^4b Taken together to form ═ O;

R ⁵ is selected from

Q is selected from C (═ O), C (═ NR) ⁸ )、NR ⁹ C(＝O)、S(＝O) ₂ Or NR ⁹ S(＝O) ₂ ；

R ⁸ 、R ⁹ Independently selected from H, C ₁ -C ₆ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7 membered heterocyclyl, C ₆ -C ₁₀ Aryl or 5-10 membered heteroaryl;

when R is ⁵ Is selected from

When R is ⁶ 、R ^7A 、R ^7B Each independently selected from H, F, Cl, Br, I, CN, carboxyl or optionally substituted by R ^6a Substituted of the following groups: c ₁ -C ₆ Alkyl radical, C ₃ -C ₁₀ Cycloalkyl, 4-10 membered heterocyclyl, or R ⁶ 、R ^7A Together with the carbon atoms to which they are attached form partially saturated C ₅ -C ₁₀ Cycloalkyl or 5-10 membered heterocyclyl, said partially saturated C ₅ -C ₁₀ Cycloalkyl or 5-10 membered heterocyclyl is optionally substituted by R ^6a Substitution;

when R is ⁵ Is selected from

When R is ^7C Selected from H or optionally substituted by R ^6a Substituted of the following groups: c ₁ -C ₆ Alkyl radical, C ₃ -C ₁₀ Cycloalkyl, 4-10 membered heterocyclyl;

R ¹⁰ is selected from C ₁ -C ₃ An alkyl group;

n is selected from 0, 1,2 or 3;

l is selected from O, NH and CH ₂ 、C(＝O)、S(＝O) ₂ Or S (═ O);

R ¹¹ selected from the group consisting of ^11a Substituted of the following groups: c ₆ -C ₁₀ Aryl, 5-10 membered heteroaryl;

each R ^1a 、R ^2a 、R ^6a 、R ^11a Independently selected from F, Cl, Br, I, OH, CN, NH ₂ O or optionally R ^b Substituted of the following groups: c ₁ -C ₆ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-10 membered heterocyclyl, C ₁ -C ₆ Alkoxy radical, C ₃ -C ₆ Cycloalkyloxy, 4-10 membered heterocyclyloxy, NH (C) ₁ -C ₆ Alkyl), S (O) ₂ (C ₁ -C ₆ Alkyl groups);

each R ^b Independently selected from F, Cl, Br, I, OH, CN, ═ O, NH ₂ 、SH、C ₁ -C ₆ Alkyl radical, C ₃ -C ₆ Cycloalkyl or 4-7 membered heterocyclyl.

In some embodiments, R ¹ Selected from the group consisting of ^1a Substituted of the following groups: phenyl, 5-10 membered heteroaryl.

In some embodiments, R ¹ Is selected from optionally substituted with R ^1a Substituted of the following groups: phenyl, 5-6 membered heteroaryl or 9-10 membered heteroaryl.

In some embodiments, R ¹ Selected from the group consisting of ^1a Substituted of the following groups: phenyl, pyridyl, benzothiazolyl.

In some embodiments, R ¹ Selected from the group consisting of ^1a A substituted phenyl group.

In some embodiments, R ^1a Selected from F, Cl, Br, I, OH, CN, NH ₂ O or optionally R ^b Substituted of the following groups: c ₁ -C ₃ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7A heterocyclic radical, C ₁ -C ₃ Alkoxy radical, C ₃ -C ₆ Cycloalkyloxy, 4-7 membered heterocyclyloxy, NH (C) ₁ -C ₃ Alkyl groups).

In some embodiments, R ^1a Selected from F, Cl, Br, I, OH, CN, NH ₂ Or the following optionally substituted with F: c ₁ -C ₃ Alkyl radical, C ₁ -C ₃ An alkoxy group.

In some embodiments, R ^1a Selected from F, Cl, NH ₂ OH, methyl or trifluoromethyl.

In some embodiments, R ¹ Is selected from

In some embodiments, R ^1a Selected from F or OH.

In some embodiments, R ¹ Is selected from

In some embodiments, R ² 、R ³ Independently selected from H, F, Cl, Br, I, CN, OH or optionally substituted with R ^2a Substituted of the following groups: NH (NH) ₂ 、C ₁ -C ₃ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7 membered heterocyclyl, C ₁ -C ₃ Alkoxy radical, C ₂ -C ₃ Alkenyl radical, C ₂ -C ₃ Alkynyl.

In some embodiments, R ² 、R ³ Independently selected from H, F, Cl, Br, I or optionally R ^2a Substituted of the following groups: c ₁ -C ₃ Alkyl radical, C ₃ -C ₆ Cycloalkyl radical, C ₁ -C ₃ Alkoxy radical, C ₂ -C ₃ Alkenyl radical, C ₂ -C ₃ Alkynyl.

In some embodiments, R ^2a Selected from halogenElement, CN, OH, ═ O or C ₁ -C ₃ An alkyl group.

In some embodiments, R ² Selected from H, halogen, ethynyl, 1-propynyl, cyclopropyl, trifluoromethoxy or difluoromethoxy.

In some embodiments, R ² Selected from H or halogen.

In some embodiments, R ² Selected from H or F.

In some embodiments, R ³ Selected from H, halogen, C ₁ -C ₃ Alkyl or C ₃ -C ₆ A cycloalkyl group.

In some embodiments, R ³ Selected from H or halogen.

In some embodiments, R ³ Selected from H, F or Cl.

In some embodiments, R ⁵ Is selected from

In some embodiments, R ⁵ Is selected from

In some embodiments, Q is selected from C (═ O) or S (═ O) ₂ 。

In some embodiments, R ⁶ 、R ^7A 、R ^7B 、R ^7C Each independently selected from H, F, Cl, Br, I, CN or optionally substituted by R ^6a Substituted of the following groups: c ₁ -C ₃ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7 membered heterocyclyl.

In some embodiments, R ^6a Selected from halogen, CN, OH, ═ O or C ₁ -C ₃ An alkyl group.

In some embodiments, R ⁶ 、R ^7A 、R ^7B 、R ^7C Each independently selected from H, F, Cl, Br, I, CN, C ₁ -C ₃ An alkyl group.

In some embodiments, R ⁶ 、R ^7A 、R ^7B 、R ^7C Each independently selected from H, F, Cl, Br, I.

In some embodiments, R ⁶ 、R ^7A 、R ^7B 、R ^7C Each independently selected from H, F.

In some embodiments, R ^7A 、R ^7B At least one of (a) is selected from H.

In some embodiments, R ^7A Selected from H, R ^7B Selected from H, F, Cl, Br, I, CN or C ₁ -C ₃ An alkyl group.

In some embodiments, R ⁶ 、R ^7A 、R ^7B 、R ^7C Each independently selected from H.

In some embodiments, R ⁵ Is selected from

In some embodiments, R ¹⁰ Selected from methyl.

In some embodiments, n is selected from 0.

In some embodiments, L is selected from O or NH.

In some embodiments, R ¹¹ Is selected from optionally substituted with R ^11a Substituted of the following groups: phenyl, 5-10 membered heteroaryl.

In some embodiments, R ¹¹ Selected from the group consisting of ^11a Substituted of the following groups: phenyl, 5-6 membered heteroaryl or 9-10 membered heteroaryl.

In some embodiments, R ¹¹ Selected from the group consisting of ^11a Substituted of the following groups: phenyl, pyrrolyl, furanyl, thienyl, pyrazolyl, thiazolyl, oxazolyl, imidazolyl, isothiazolyl, isoxazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzopyrolyl, benzofuranyl, benzothienyl, benzopyrazolyl, pyrimidazolyl, pyridopyrazolyl, benzopyridyl, benzopyrimidinyl, benzopyrazinyl, and benzopyrazinyl.

In some embodiments, R ¹¹ Selected from the group consisting of ^11a Substituted of the following groups: phenyl, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl.

In some embodiments, R ¹¹ Selected from the group consisting of ^11a Substituted of the following groups: phenyl, pyridyl.

In some embodiments, R ^11a Selected from F, Cl, Br, I, OH, CN, NH ₂ O or optionally R ^b Substituted of the following groups: c ₁ -C ₆ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7 membered heterocyclyl, C ₁ -C ₆ An alkoxy group.

In some embodiments, R ^11a Is selected from C ₁ -C ₃ An alkyl group.

In some embodiments, R ^11a Selected from methyl or isopropyl.

In some embodiments, R ¹¹ Is selected from

In some embodiments, the compound of formula (I), or a pharmaceutically acceptable salt thereof, is selected from a compound of formula (II), or a pharmaceutically acceptable salt thereof:

wherein R is ¹ 、R ² 、R ³ 、R ^4a 、R ^4b 、R ¹¹ L is as defined above.

In some embodiments, the compound of formula (I), or a pharmaceutically acceptable salt thereof, is selected from the following compounds, or a pharmaceutically acceptable salt thereof:

in some embodiments, the compound of formula (I), or a pharmaceutically acceptable salt thereof, is selected from the following compounds, or a pharmaceutically acceptable salt thereof:

in some embodiments, the compound of formula (I), or a pharmaceutically acceptable salt thereof, is selected from the following compounds, or a pharmaceutically acceptable salt thereof:

the invention also provides a pharmaceutical composition which comprises the compound shown in the formula (I) or pharmaceutically acceptable salt thereof and pharmaceutically acceptable auxiliary materials.

Further, the invention relates to an application of the compound shown in the formula (I) or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof in preparing a medicament for preventing or treating KRAS G12C related diseases.

Further, the invention relates to application of the compound shown in the formula (I) or pharmaceutically acceptable salts thereof or a pharmaceutical composition thereof in preventing or treating KRAS G12C related diseases.

Further, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for preventing or treating a KRAS G12C-related disease.

The invention also relates to a method of treating a disease associated with KRAS G12C, the method comprising administering to a patient a therapeutically effective amount of a pharmaceutical formulation comprising a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof.

Preferred embodiments of the present invention wherein the KRAS G12C-associated diseases include, but are not limited to, inflammatory diseases, autoimmune diseases, and cancer.

Definition and description of terms

Unless otherwise indicated, the definitions of groups and terms described in the specification and claims of the present invention, including definitions thereof as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. The definitions of the groups and the structures of the compounds in such combinations and after the combination are within the scope of the present invention as defined in the specification.

The term "pharmaceutically acceptable salts" refers to pharmaceutically acceptable salts of non-toxic acids or bases, including salts of inorganic acids and bases, organic acids and bases.

The term "stereoisomer" refers to isomers resulting from the different arrangement of atoms in a molecule, including cis-trans isomers, enantiomers, and diastereomers.

The compounds of the present invention may have an asymmetric atom such as a carbon atom, a sulfur atom, a nitrogen atom, a phosphorus atom (optical center) or an asymmetric double bond. Racemates, enantiomers, diastereomers, geometric isomers are included within the scope of the present invention.

The enantiomers or enantiomerically pure compounds herein are illustrated by Maehr, J.chem.Ed.1985, 62: 114-120. Using wedge and virtual wedge keys, unless otherwise indicated (

And

) Representing the absolute configuration of a stereocenter, using black solid and imaginary bonds (

And

) Represents a cis-trans configuration of an alicyclic compound. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, they include the E, Z geometric isomer unless otherwise specified. Likewise, all tautomeric forms are included within the scope of the invention.

The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-) -and (+) -enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, as well as racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. An additional asymmetric carbon atom, asymmetric sulfur atom, asymmetric nitrogen atom or asymmetric phosphorus atom may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are included within the scope of the present invention. The compounds of the present application containing asymmetric atoms can be isolated in optically active pure form or in racemic form. The optically active pure form can be resolved from a racemic mixture or synthesized by using chiral starting materials or chiral reagents.

The term "tautomer" refers to an isomer of a functional group resulting from the rapid movement of an atom in two positions in a molecule. The compounds of the invention may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer often result in a mixture whose physicochemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; whereas in phenol the enol type predominates. The present invention encompasses all tautomeric forms of the compounds.

The term "pharmaceutical composition" denotes a mixture of one or more compounds described herein or a physiologically/pharmaceutically acceptable salt or prodrug thereof with other chemical components, e.g. physiologically/pharmaceutically acceptable excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to an organism.

The term "substituted" means that any one or more hydrogen atoms on a particular atom is replaced with a substituent, so long as the valence of the particular atom is normal and the substituted compound is stable. When the substituent is oxo (i.e., ═ O), meaning that two hydrogen atoms are substituted, oxo does not occur on the aryl.

The terms "optionally" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not. For example, ethyl is "optionally" substituted with halo, meaning that ethyl may be unsubstituted (CH) ₂ CH ₃ ) Monosubstituted (e.g. CH) ₂ CH ₂ F) Polysubstituted (e.g. CHFCH) ₂ F、CH ₂ CHF ₂ Etc.) or completely substituted (CF) ₂ CF ₃ ). It will be appreciated by those skilled in the art that any group containing one or more substituents will not incorporate any substitution or substitution pattern which is sterically impossible and/or cannot be synthesized.

The term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

The term "C ₅ -C ₁₀ Cycloalkyl "is to be understood as meaning saturated or partially saturated, nonaromatic, monovalent cycloalkyl radicals having 5,6,7,8, 9 or 10 carbon atoms.

The term "C ₁ -C ₆ Alkyl "is understood to mean a straight-chain or branched, saturated monovalent hydrocarbon radical having 1,2,3,4, 5 or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-ethylpropyl, 1, 2-dimethylpropyl, neopentyl, 1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 2, 3-dimethylbutyl, 2-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 2, 3-dimethylbutyl, 2-methylpentyl, 1-methyl-pentyl, 1-ethylpropyl, 1-dimethylpropyl, neopentyl, or neopentyl or an amino group, or a corresponding to be mentioned,1, 3-dimethylbutyl or 1, 2-dimethylbutyl, etc.; preferably, "C ₁ -C ₆ Alkyl groups "may contain" C ₁ -C ₃ Alkyl group and C ₁ -C ₃ Alkyl "is understood to mean a straight-chain or branched saturated monovalent hydrocarbon radical having 1,2,3 carbon atoms.

The term "alkoxy" is understood to mean "alkyloxy" or "alkyl-O-", preferably "C ₁ -C ₆ Alkoxy groups "may contain" C ₁ -C ₃ Alkoxy ".

The term "C ₃ -C ₁₀ Cycloalkyl "is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 10 carbon atoms. Such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group such as a decaline ring. Preferably, "C ₃ -C ₁₀ Cycloalkyl groups "may comprise" C ₃ -C ₆ Cycloalkyl radical ", the term" C ₃ -C ₆ Cycloalkyl "is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms.

The term "cycloalkyloxy" is understood to mean "cycloalkyl-O-", preferably "C" ₃ -C ₁₀ Cycloalkyloxy "may contain" C ₃ -C ₆ Cycloalkyloxy ".

The term "4-10 membered heterocyclyl" means a saturated or partially saturated monovalent monocyclic, fused, spiro or bridged ring containing 1-5, preferably 1-3 heteroatoms selected from N, O, B and S. In particular, "4-10 membered heterocyclic group" includes "5-10 membered heterocyclic group", "4-7 membered heterocyclic group"; the "4-10 membered heterocyclic group" includes, but is not limited to: 4-membered rings such as azetidinyl, oxetanyl; a 5-membered ring such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 1,3, 2-dioxaboropentyl; or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl; or a partially saturated 6-membered ring such as tetrahydropyridinyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclyl group may be bicyclic, for example but not limited to a 5,5 membered ring, such as a hexahydrocyclopenta [ c ] pyrrol-2 (1H) -yl ring, or a 5,6 membered bicyclic ring, such as a hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl ring. The nitrogen atom containing ring may be partially unsaturated, i.e., it may contain one or more double bonds, such as, but not limited to, 2, 5-dihydro-1H-pyrrolyl, 4H- [1,3,4] thiadiazinyl, 4, 5-dihydrooxazolyl, or 4H- [1,4] thiazinyl, or it may be benzo-fused, such as, but not limited to, dihydroisoquinolinyl. Optionally, the 4-10 membered heterocyclyl may be "4-10 membered heterocycloalkyl," meaning a saturated, monovalent monocyclic, fused, spiro, or bridged ring containing 1-5 heteroatoms; preferably, "4-10 membered heterocycloalkyl" includes 5-10 membered heterocycloalkyl and 4-7 membered heterocycloalkyl, also including 5-6 membered heterocycloalkyl and the like.

The term "heterocyclyloxy" is understood to mean "heterocyclyl-O-".

The term "C ₆ -C ₁₀ Aryl "is understood to preferably mean a mono-, bi-or tricyclic hydrocarbon ring having a monovalent or partially aromatic character with 6 to 10 carbon atoms. In particular a ring having 6 carbon atoms ("C) ₆ Aryl "), such as phenyl; or a ring having 9 carbon atoms ("C) ₉ Aryl), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C) ₁₀ Aryl), such as tetralinyl, dihydronaphthyl, or naphthyl.

The term "5-10 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic aromatic ring systems having 5 to 10 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, including but not limited to thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl and the like and their benzo derivatives, such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl and the like; or pyridyl, piperazinyl, pyrimidinyl, pyrazinyl, triazinyl, and the like, and benzo derivatives thereof, such as quinolinyl, quinazolinyl, isoquinolinyl, and the like. "5-6 membered heteroaryl" refers to a monovalent monocyclic aromatic ring system having 5 or 6 ring atoms and which contains 1-4, preferably 1-3 heteroatoms independently selected from N, O and S. "9-10 membered heteroaryl" refers to a monovalent bicyclic aromatic ring system having 9 or 10 ring atoms and which contains 1-5, preferably 1-3 heteroatoms independently selected from N, O and S.

The term "C ₂ -C ₆ Alkenyl "is understood to mean a straight-chain or branched monovalent hydrocarbon radical which contains one or more double bonds and has 2,3,4, 5 or 6 carbon atoms, examples including, but not limited to, vinyl (-CH ═ CH) ₂ ) Prop-1-enyl (-CH ═ CHCH) ₃ ) Prop-2-enyl (-CH) ₂ CH＝CH ₂ ) 2-methylpropan-1-enyl, but-2-enyl, but-3-enyl, but-1, 3-dienyl, 2-methyl-1, 3-butadienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl or hex-4-enyl. Preferably, "C ₂ -C ₆ Alkenyl "includes" C ₂ -C ₃ Alkenyl ".

The term "C ₂ -C ₆ Alkynyl is understood to mean a straight-chain or branched monovalent hydrocarbon radical which contains one or more triple bonds and has 2,3,4, 5 or 6 carbon atoms, examples including, but not limited to, ethynyl (-C.ident.CH), prop-1-ynyl (1-propynyl, -C.ident.CCH) ₃ ) Prop-2-ynyl (propargyl), but-1-ynyl, but-2-ynyl or but-3-ynyl. "C ₂ -C ₃ Examples of alkynyl include ethynyl (-C.ident.CH), prop-1-ynyl (1-propynyl, -C.ident.CCH) ₃ ) Prop-2-ynyl (propargyl).

In particular, the term "treating" means administering a compound or formulation described herein to prevent, ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes:

(i) preventing the occurrence of a disease or condition in a mammal, particularly when such mammal is susceptible to the disease condition, but has not yet been diagnosed as having the disease condition;

(ii) inhibiting the disease or disease state, i.e., arresting its development;

(iii) alleviating the disease or condition, i.e., causing regression of the disease or condition.

The term "therapeutically effective amount" means an amount of a compound of the invention that (i) treats or prevents a particular disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of a compound of the present invention that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by those skilled in the art with their own knowledge and this disclosure.

The term "adjuvant" refers to a pharmaceutically acceptable inert ingredient. Examples of classes of the term "excipient" include, without limitation, binders, disintegrants, lubricants, glidants, stabilizers, fillers, diluents, and the like. Excipients enhance the handling characteristics of the pharmaceutical formulation, i.e., make the formulation more amenable to direct compression by increasing flowability and/or cohesiveness. Examples of typical "pharmaceutically acceptable carriers" suitable for use in the above formulations are: saccharides, starches, cellulose and its derivatives and the like are commonly used as excipients in pharmaceutical preparations.

The term "pharmaceutically acceptable adjuvants" refers to those adjuvants which do not have a significant irritating effect on the organism and do not impair the biological activity and properties of the active compound. Suitable adjuvants are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and/or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

The words "comprise", "comprises", "comprising" or "including" and variations thereof such as "comprises" or "comprising" are to be interpreted in an open, non-exclusive sense, i.e., "including but not limited to".

The compounds of the present invention may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combinations thereof with other chemical synthetic methods, and equivalents thereof known to those skilled in the art, with preferred embodiments including, but not limited to, examples of the present invention.

The present application also includes isotopically-labeled compounds of the present application, which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine and chlorine, such as respectively ² H、 ³ H、 ¹¹ C、 ¹³ C、 ¹⁴ C、 ¹³ N、 ¹⁵ N、 ¹⁵ O、 ¹⁷ O、 ¹⁸ O、 ³¹ P、 ³² P、 ³⁵ S、 ¹⁸ F、 ¹²³ I、 ¹²⁵ I and ³⁶ cl, and the like.

Certain isotopically-labelled compounds of the present application (e.g. with ³ H and ¹⁴ c-labeled ones) can be used in compound and/or substrate tissue distribution assays. Tritiated (i.e. by tritiation) ³ H) And carbon-14 (i.e. ¹⁴ C) Isotopes are particularly preferred for their ease of preparation and detectability. Positron emitting isotopes, such as ¹⁵ O、 ¹³ N、 ¹¹ C and ¹⁸ f can be used in Positron Emission Tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the present application can generally be prepared by following procedures analogous to those disclosed in the schemes and/or in the examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

In addition, heavier isotopes are used (such as deuterium (i.e., deuterium) ² H) Substitution may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements), and thus may be preferred in certain circumstances where deuterium substitution may be partial or complete, partial deuterium substitution meaning that at least one hydrogen is substituted with at least one deuterium.

The pharmaceutical compositions of the present application can be prepared by combining the compounds of the present application with suitable pharmaceutically acceptable excipients, for example, can be formulated into solid, semi-solid, liquid or gaseous formulations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres, aerosols, and the like.

Typical routes of administration of a compound of the present application or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.

The pharmaceutical compositions of the present application can be manufactured by methods well known in the art, such as conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, lyophilizing, and the like.

In some embodiments, the pharmaceutical composition is in an oral form. For oral administration, the pharmaceutical compositions may be formulated by mixing the active compounds with pharmaceutically acceptable excipients well known in the art. These adjuvants enable the compounds of the present application to be formulated as tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions and the like, for oral administration to a patient.

Solid oral compositions may be prepared by conventional mixing, filling or tableting methods. For example, it can be obtained by the following method: the active compounds are mixed with solid adjuvants, optionally the mixture obtained is milled, if desired with further suitable adjuvants, and the mixture is then processed to granules, to give tablets or dragee cores. Suitable excipients include, but are not limited to: binders, diluents, disintegrants, lubricants, glidants, sweeteners or flavoring agents and the like.

The pharmaceutical compositions may also be adapted for parenteral administration, as sterile solutions, suspensions or lyophilized products in suitable unit dosage forms.

In all methods of administration of the compounds of the general formula I described herein, the daily dose is from 0.01 to 100mg/kg body weight, preferably from 0.05 to 50mg/kg body weight, more preferably from 0.1 to 30mg/kg body weight, in single or divided doses.

The chemical reactions of the embodiments of the present invention are carried out in a suitable solvent that is compatible with the chemical changes of the present invention and the reagents and materials required therefor. In order to obtain the compounds of the present invention, it is sometimes necessary for a person skilled in the art to modify or select the synthesis steps or reaction schemes based on the existing embodiments.

Detailed Description

The following examples illustrate the technical solutions of the present invention in detail, but the scope of the present invention includes but is not limited thereto.

Unless otherwise specified, the ratios expressed by the mixed solvents are volume mixing ratios.

Unless otherwise specified,% means weight percent wt%.

The compounds are prepared by hand or

The software names, and the commercial compounds are under the supplier catalog name.

The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) and/or Mass Spectrometry (MS). NMR shift in units of 10 ^-6 (ppm). Solvents for NMR determination are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol and the like, and an internal standard is Tetramethylsilane (TMS); ' IC ₅₀ "half inhibitory concentration" means the concentration at which half of the maximum inhibitory effect is achieved. The following eluent can be a mixed eluent formed by two or more solvents, wherein the ratio is the volume ratio of each solvent, for example, "0-10% methanol/dichloromethane" represents methanol in the mixed eluent during gradient elution: the volume consumption of the dichloromethane is 0: 100-10: 100.

Preparation example 1 Synthesis of 2-isopropyl-4-methylpyridin-3-ol

To a solution of 2-isopropyl-4-methylpyridin-3-amine (4g,26.63mmol) in sulfuric acid (6.40g,65.25mmol) and water (30mL) at 0 deg.C was added dropwise an aqueous solution (18mL) of sodium nitrite (2.39g,34.62 mmol). The reaction was stirred at 0 ℃ for 30min, then at 25 ℃ for 16 h. After the reaction was completed, saturated aqueous sodium bicarbonate was added to adjust the pH to 7. The product is prepared by using a mixed solvent of chloroform: isopropanol 3:1(100mL × 3), the organic phase was dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent to give 2-isopropyl-4-methylpyridin-3-ol (4 g).

MS m/z(ESI):＝152.2[M+H] ⁺ 。

Example 1- ((12aS) -10-chloro-9- (2-fluoro-6-hydroxyphenyl) -7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -3,4,12,12 a-tetrahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepin-2 (1H) -yl) prop-2-en-1-one (Compound 1)

Step 1 Synthesis of methyl 2-fluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxy ] -3-nitrobenzoate (intermediate 1-2)

Reaction 1-1(6.7g,30.86mmol) was dissolved in DMF (30ml), sodium hydride (2.47g,61.72mmol, effective content 60%) was added at 0 ℃ and stirred at 0 ℃ for 30 minutes, 2-isopropyl-4-methylpyridin-3-ol (5.2g,34.39mmol) was dissolved in DMF (10ml), added dropwise to the reaction flask at 0 ℃ and stirred at 0 ℃ for 30 minutes. LCMS showed the starting material reaction was complete. The reaction mixture was poured into a saturated ammonium chloride solution (100ml), extracted with ethyl acetate (100 ml. times.2), washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by flash column on silica gel (

80g

Flash column on silica gel eluting with a 10-20% ethyl acetate/petroleum ether gradient @60mL/min) afforded the title compound (6 g).

MS m/z(ESI):＝349[M+H] ⁺

Step 2 Synthesis of methyl 3-amino-2-fluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxy ] benzoate (intermediate 1-3)

Intermediate 1-2(2g,5.74mmol) was dissolved in methanol (30ml) and Pd/C (30.55mg, 287.09. mu. mol) was added and stirred under hydrogen (15psi) at 25 ℃ for 16 h. LCMS showed the reaction was complete. The reaction solution was filtered through celite, followed by concentration under reduced pressure. Rapid passage of residueSilica gel column purification (

20g

Flash column of silica gel eluting with 30-40% ethyl acetate/petroleum ether gradient @18 ml/min). The title compound (800mg) was obtained.

MS m/z(ESI):＝319[M+H] ⁺

Step 3 Synthesis of methyl 3-amino-4-bromo-2-fluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxy ] benzoate (intermediate 1-4)

After dissolving intermediate 1-3(4.6g,14.45mmol) in acetic acid (40mL), pyridinium tribromide (6.01g,18.78mmol) was added in portions, and the reaction mixture was stirred at 45 ℃ for 5 hours. LCMS showed the reaction was complete. The reaction solution was poured into 20 wt% sodium sulfite solution, stirred for 10 minutes, extracted with ethyl acetate (10mL × 3), washed with saturated brine, and the organic phase was concentrated under reduced pressure. The residue was purified by flash column on silica gel (

40g

Flash silica gel column, eluent 30-40% ethyl acetate/petroleum ether gradient @36 ml/min). The title compound (4.8g) was obtained.

MS m/z(ESI):＝399[M+H] ⁺

Step 4 Synthesis of methyl 4-bromo-3-chloro-2-fluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxy ] benzoate (intermediate 1-5)

Copper chloride (3.25g,24.17mmol) and tert-butyl nitrite (3.12g,30.21mmol) were dissolved in acetonitrile (70mL) and stirred for 10 min, intermediates 1-4(4.8g,12.08mmol) were dissolved in acetonitrile (50mL), added dropwise to the reaction flask at-20 deg.C, then warmed to 20 deg.C and stirred at 20 deg.C for 16 h. LCMS showed the starting material reacted, the reaction was poured into 6M aqueous hydrochloric acid (60mL), extracted with ethyl acetate (100mL × 3), dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by flash column on silica gel (

40g

Flash column of silica gel, eluent 10% methanol/dichloromethane gradient @100 mL/min.) gave the title compound (4.9 g).

MS m/z(ESI):＝417.8[M+H] ⁺

Step 5 Synthesis of 4-bromo-3-chloro-2-fluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxy ] benzoic acid (intermediate 1-6)

Intermediate 1-5(500mg,1.20mmol) was dissolved in tetrahydrofuran (5mL) and water (5mL), lithium hydroxide (287.40mg,12.00mmol) was added, and the mixture was stirred at 60 ℃ for 16 hours. LCMS showed the reaction was complete. The reaction solution was cooled to 25 ℃ and water (20mL) was added, followed by extraction with ethyl acetate (20 mL). The aqueous phase was taken out, the pH was adjusted to 1-3 with 1M aqueous hydrochloric acid, extraction was performed with isopropanol/dichloromethane 1:3, and the organic phase was taken out, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to give the title compound (300 mg).

MS m/z(ESI):＝403.8[M+H] ⁺

Step 6 Synthesis of (S) -4- [ 4-bromo-3-chloro-2-fluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxy ] benzoyl ] -3- (hydroxymethyl) piperazine-1-carboxylic acid tert-butyl ester (intermediate 1-7)

Intermediate 1-6(300mg, 745.08. mu. mol) was dissolved in dichloromethane (10mL), oxalyl chloride (283.71mg,2.24mmol) was added, the mixture was stirred at 0 ℃ for 20 minutes, LCMS indicated complete conversion of the starting material, the reaction was concentrated under reduced pressure, and the residue was dissolved in dichloromethane (10 mL). A methylene chloride solution of the residue was added dropwise to a methylene chloride solution containing tert-butyl (S) -3- (hydroxymethyl) piperazine-1-carboxylate (193.37mg, 894.09. mu. mol) and triethylamine (301.58mg,2.98mmol) at 0 ℃. After dropping, the temperature was gradually increased and the mixture was stirred at 20 ℃ for 3 hours. LCMS showed the reaction was complete. After quenching with water (15mL), extraction with dichloromethane (20mL × 3) and washing with saturated brine, drying over anhydrous magnesium sulfate and concentration under reduced pressure, the title compound was obtained as a crude product (480 mg).

MS m/z(ESI):＝602[M+H] ⁺

Step 7 Synthesis of (S) -9-bromo-10-chloro-7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -6-oxo-3, 4,12,12 a-tetrahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepine-2 (1H) -carboxylic acid tert-butyl ester (intermediate 1-8)

Intermediate 1-7(450mg, 748.87. mu. mol) was dissolved in DMF (12mL), and sodium hydrogen (89.86mg,2.25mmol, effective content 60%) was added at 0 ℃ to stir the reaction solution at 0 ℃ for 20 minutes. LCMS showed the starting material was reacted. The reaction mixture was poured into an aqueous solution of ammonium chloride (30mL), extracted with ethyl acetate (20mL × 3), washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by flash column on silica gel (

4g

Flash column of silica gel eluting with 20-33% ethyl acetate/petroleum ether gradient @18 mL/min). The title compound (220mg) was obtained.

MS m/z(ESI):＝582[M+H] ⁺

Step 8 Synthesis of (S) -9-bromo-10-chloro-7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -3,4,12,12 a-tetrahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepine-2 (1H) -carboxylic acid tert-butyl ester (intermediate 1-9)

Intermediate 1-8(220.00mg, 378.72. mu. mol) was dissolved in tetrahydrofuran (15mL), borane tetrahydrofuran (1M,7.57mL) was added at 0 ℃ and the reaction was heated to 60 ℃ and stirred for 16 hours. LCMS showed the reaction was complete. Methanol (15mL) was added slowly to the reaction at 0 deg.C, warmed to 60 deg.C, stirred for 1 hour, then cooled to 25 deg.C, and concentrated under reduced pressure. The residue was purified by thin layer preparative chromatography (silica, petroleum ether: ethyl acetate 4: 5). The title compound (180mg) was obtained.

MS m/z(ESI):＝568[M+H] ⁺

Step 9 Synthesis of (12aS) -10-chloro-9- (2-fluoro-6-hydroxyphenyl) -7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -3,4,12,12 a-tetrahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepine-2 (1H) -carboxylic acid tert-butyl ester (intermediate 1-10)

After dissolving intermediate 1-9(180mg, 317.51. mu. mol) and 2-fluoro-6-hydroxyphenyltrifluoroborate potassium salt (346.08mg,1.59mmol) in toluene (6mL), potassium carbonate (131.64mg, 952.53. mu. mol) and (2-dicyclohexylphosphino-2, 6-dimethoxybiphenyl) [2- (2-amino-1, 1-biphenylyl) ] palladium (II) methanesulfonate (SPhos Pd G3) (24.77mg, 31.75. mu. mol), nitrogen gas were added, followed by heating to 100 ℃ and stirring for 16 hours. LCMS showed the reaction was complete. The reaction mixture was filtered through celite, extracted with ethyl acetate (10mL), washed with water (10mL), and concentrated to dryness under reduced pressure. The residue was purified twice by thin layer preparative chromatography (silica, ethyl acetate: petroleum ether ═ 2: 1). The title compound (30mg) was obtained.

MS m/z(ESI):＝598[M+H] ⁺

Step 10 Synthesis of 2- ((S) -10-chloro-7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -1,2,3,4,12,12 a-hexahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepin-9-yl) -3-fluorophenol (intermediate 1-11)

Intermediate 1-10 was dissolved in dichloromethane (1mL), trifluoroacetic acid (770.00mg,6.75 mmol) was added and the mixture was stirred at 25 ℃ for 2 h. TLC (petroleum ether: ethyl acetate 1:1) monitored completion of the reaction, and the reaction solution was concentrated to dryness under reduced pressure to give the crude title compound (31 mg).

MS m/z(ESI):＝499[M+H] ⁺

Step 11 Synthesis of 1- ((12aS) -10-chloro-9- (2-fluoro-6-hydroxyphenyl) -7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -3,4,12,12 a-tetrahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepin-2 (1H) -yl) prop-2-en-1-one (Compound 1)

Intermediate 1-11(31mg,50.65 μmol) was dissolved in dichloromethane (2mL), diisopropylethylamine (19.64mg,151.96 μmol) was added to adjust pH 8, acryloyl chloride (3.90mg,43.05 μmol) was dissolved in dichloromethane (0.2mL) at 0 ℃ and the solution was added dropwise to the reaction flask. The reaction solution was stirred at 16 ℃ for 16 hours. LCMS showed the reaction was complete. The reaction solution was concentrated to dryness under reduced pressure. The residue was purified by preparative high performance liquid chromatography (basic conditions, column: Boston Prime C18150 x 30mM x 5 um; mobile phase: A: water (0.04% aqueous ammonia +10mM ammonium bicarbonate), B: acetonitrile ]; B%: 43% -65%, 7 min) to give one pair of isomers (atropisomers) Compound 1 isomer 1(1.8mg, LCMS retention time: 2.286min), Compound 1 isomer 2(1.0mg, LCMS retention time: 2.445 min).

LCMS detection conditions: waters Xbridge C1830 × 2.0mm,3.5 um; mobile phase a) 0.04% aqueous solution of trifluoroacetic acid; B) 0.02% trifluoroacetic acid in acetonitrile; gradient elution, increasing mobile phase B from 0% to 95% in 5.8 min; mobile phase B95% was held for 1.1 min; mobile phase B was then reduced to 0% at 6.91 minutes and maintained for 0.09 minutes. The flow rate was 1.0mL/min

MS m/z(ESI):＝552[M+H] ⁺

Compound 1 isomer 1 ¹ H NMR:(400MHz,Methanol-d ₄ )δ＝8.25(d,J＝5.0Hz,1H),7.22-7.17(m,1H),7.16-7.08(m,1H),6.89-6.71(m,1H),6.61(t,J＝7.4Hz,1H),6.55(t,J＝8.8Hz,1H),6.24(d,J＝16.1Hz,1H),5.86(d,J＝3.8Hz,1H),5.78(d,J＝10.0Hz,1H),4.54-4.41(m,2H),4.31-4.29(m,0.5H),4.25-4.11(m,0.5H),4.00(d,J＝11.5Hz,1H),3.93-3.78(m,2H),3.60-3.47(m,0.5H),3.38-3.33(m,0.5H),3.28-3.12(m,1.5H),3.06-3.00(m,1.5H),2.95(s,1H),2.66(s,1H),2.21(s,1.5H),2.15(s,1.5H),1.30-1.19(m,3H),1.16-1.09(m,3H).

Compound 1 isomer 2 ¹ H NMR:(400MHz,Methanol-d ₄ )δ8.25(d,J＝5.0Hz,1H),7.19(t,J＝5.1Hz,1H),7.16-7.08(m,1H),6.89-6.74(m,1H),6.65-6.59(m,1H),6.55(t,J＝8.7Hz,1H),6.24(d,J＝15.8Hz,1H),5.87(d,J＝11.5Hz,1H),5.78(t,J＝8.7Hz,1H),4.49-4.40(m,1H),4.45-4.40(m,1H),4.22-4.20(m,0.5H),4.16-4.06(m,0.5H),4.01-3.82(m,3H),3.58-3.55(m,0.5H),3.40-3.30(m,1H),3.28-3.22(m,0.5H),3.19-3.11(m,1H),3.07(d,J＝12.5Hz,1H),3.00(s,1H),2.67(s,1H),2.24(s,1.5H),2.12(s,1.5H),1.27-1.05(m,6H).

Example 2,1- ((12aS) -10-chloro-8-fluoro-9- (2-fluoro-6-hydroxyphenyl) -7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -3,4,12,12 a-tetrahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepin-2 (1H) -yl) prop-2-en-1-one (Compound 2)

Step 1 Synthesis of methyl 2, 5-difluoro-6- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -3-nitrobenzoate (intermediate 2-2)

To a solution of reactant 2-1(3.60g,15.32mmol) and 2-isopropyl-4-methylpyridin-3-ol (2.2g,14.55mmol) in DMF (5mL) was added sodium hydride (1.04g,26.04mmol, 60% available content) with stirring at 0 deg.C, the reaction was stirred at 0 deg.C for 30min and LCMS indicated complete reaction of the starting material. After completion of the reaction, the reaction mixture was poured into a saturated ammonium chloride solution (100mL), extracted with ethyl acetate (100 mL. x.2), washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated to dryness under reduced pressure. The residue was purified by flash column chromatography (

40g

Flash silica gel column, eluent 10-20% ethyl acetate/petroleum ether gradient @40 mL/min). The title compound (4g) was obtained.

MS m/z(ESI):＝367[M+H] ⁺

Step 2 Synthesis of methyl 3-amino-2, 5-difluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxy ] benzoate (intermediate 2-3)

Intermediate 2-2(4g,10.92mmol) was dissolved in methanol (100mL), wet palladium on carbon (1.16g,545.98 μmol, 5% active content) was added and stirred under hydrogen atmosphere at 20 ℃ for 5 h. LCMS showed the starting material reaction was complete. The reaction solution was filtered through celite, and the filtrate was concentrated under reduced pressure to give the crude title compound (3.5 g).

MS m/z(ESI):＝337[M+H] ⁺

Step 3 Synthesis of methyl 3-amino-4-bromo-2, 5-difluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxybenzoate (intermediate 2-4)

To a solution of intermediate 2-3(3.5g,10.41mmol) in acetic acid (50mL) at 20 ℃ was added tribromopyridinium salt (3.66g,11.45mmol), warmed to 50 ℃ and stirred at 50 ℃ for 5 hours, LCMS showed reaction completion. The reaction mixture was concentrated under reduced pressure to remove acetic acid, quenched with sodium sulfite solution, adjusted to pH 8 with sodium carbonate solid, added with ethyl acetate (100mL), and extracted with ethyl acetate (100mL × 2), dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to dryness. Rapid passing of residue through siliconPurifying with a gel column (

40g

Flash column on silica gel, eluent 20-30% ethyl acetate/petroleum ether gradient @40mL/min) afforded the crude title compound (4.2 g).

MS m/z(ESI):＝415.9[M+H] ⁺

Step 4 Synthesis of methyl 4-bromo-3-chloro-2, 5-difluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxy ] benzoate (intermediate 2-5)

Copper chloride (2.66g,19.75mmol) and tert-butyl nitrite (2.55g,24.69mmol) were added to acetonitrile (40mL), stirred at 20 ℃ for 10 minutes, intermediate 2-4(4.1g,9.87mmol) was dissolved in acetonitrile (40mL) and added to the reaction flask at 0 ℃ and the reaction stirred at 25 ℃ for 16 hours. LCMS showed the starting material was reacted. The reaction mixture was poured into 1M HCl (20mL), ethyl acetate (50mL), water (30mL), concentrated under reduced pressure, adjusted to pH 8 with sodium carbonate, extracted with ethyl acetate (50mL × 3), dried over anhydrous magnesium sulfate, and concentrated to dryness under reduced pressure. The residue was purified by flash column chromatography (

40g

Flash column on silica, eluent 15% ethyl acetate/petroleum ether gradient @40mL/min) to afford the title compound (3.8 g).

MS m/z(ESI):＝435.8[M+H] ⁺

Step 5 Synthesis of 4-bromo-3-chloro-2, 5-difluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxy ] benzoic acid (intermediate 2-6)

Intermediate 2-5(3.8g,8.74mmol) was dissolved in tetrahydrofuran (20mL), water (20mL), lithium hydroxide monohydrate (5.50g,131.14mmol) was added thereto, and the reaction solution was stirred at 50 ℃ for 5 hours. LCMS showed the starting material reacted to completion and the product formed. The reaction solution was adjusted to pH 1 with 3M hydrochloric acid solution, filtered to obtain a solid, the solid was dried to obtain 1.9g, the filtrate was extracted with ethyl acetate (30mL × 3), dried over anhydrous magnesium sulfate, concentrated under reduced pressure to obtain a solid (1.9g), and the two batches of solid were combined. The title compound (3.8g, crude) was obtained.

MS m/z(ESI):＝421.8[M+H] ⁺

Step 6 Synthesis of (3S) -4- [ 4-bromo-3-chloro-2, 5-difluoro-6- [ (2-isopropyl-4-methylpyridin-3-yl) oxy ] benzoyl ] -3- (hydroxymethyl) piperazine-1-carboxylic acid tert-butyl ester (intermediate 2-8)

Oxalyl chloride (1.72g,13.55mmol) is added to a solution of intermediate 2-6(1.9g,4.52mmol) in dichloromethane (20mL) at 0 ℃, the temperature is raised to 20 ℃, stirring is carried out for 2 hours, LCMS shows that the raw materials are reacted completely, and the solvent is removed by concentration under reduced pressure to obtain intermediate 2-7; the intermediate 2 to 7 was dissolved in dichloromethane (15mL), and the reaction solution was added dropwise to a solution of tert-butyl (S) -3- (hydroxymethyl) piperazine-1-carboxylate (1.07g,4.97mmol) and triethylamine (1.83g,18.07mmol) in dichloromethane (15mL), and the reaction was stirred for 3 hours. After the reaction is finished. Water (20mL) was added to the reaction solution, extracted with dichloromethane (20mL × 3), dried over anhydrous magnesium sulfate, and concentrated to dryness under reduced pressure. The residue was purified by flash column chromatography (

40g

Flash silica gel column, eluent 15-40% ethyl acetate/petroleum ether gradient @40 mL/min). The title compound (2.07g) was obtained.

MS m/z(ESI):＝619.9[M+H] ⁺

Step 7 Synthesis of (S) -9-bromo-10-chloro-8-fluoro-7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -6-oxo-3, 4,12,12 a-tetrahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepine-2 (1H) -carboxylic acid tert-butyl ester (intermediate 2-9)

To a solution of intermediate 2-8(2g,3.23mmol) in DMF (20mL) at 0 deg.C was added sodium hydride (387.75mg,9.69mmol, 60% active content) and the reaction was stirred at 0 deg.C for 2 h. LCMS showed the starting material reaction was complete. The reaction solution was poured into ammonium chloride solution (100mL), extracted with ethyl acetate (100 mL. x.2), washed with saturated brine, and concentrated to dryness under reduced pressure. The residue was purified by flash column chromatography (

20g

Flash silica gel column, eluent 15-35% tetrahydrofuran/petroleum ether, gradient @35 mL/min). The title compound (1.0g) was obtained.

MS m/z(ESI):＝598[M+H] ⁺

Step 8 Synthesis of (12aS) -10-chloro-8-fluoro-9- (2-fluoro-6-hydroxyphenyl) -7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -6-oxo-3, 4,12,12 a-tetrahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepine-2 (1H) -carboxylic acid tert-butyl ester (intermediate 2-10)

Intermediate 2-9(250mg, 417.44. mu. mol) and potassium 2-fluoro-6-hydroxyphenyltrifluoroborate salt (91.00mg, 417.44. mu. mol) were dissolved in dioxane (6mL) and water (1.5mL), potassium carbonate (173.08mg,1.25mmol), RuPhos (19.48mg, 41.74. mu. mol) and methanesulfonic acid (2-dicyclohexylphosphine-2, 6-diisopropoxy-1, 1-biphenyl) (2-amino-1, 1-biphenyl-2-yl) palladium (II) (RuPhos Pd G3) (34.91mg, 41.74. mu. mol) were added, and the reaction was stirred at 95 ℃ for 3 hours under a nitrogen atmosphere. After the reaction, the temperature is reduced to 20 ℃, ethyl acetate (10mL) is used for extraction for three times, organic phases are combined, and the organic phase is decompressed and concentrated (0.01MPa) to remove the solvent. The residue was purified by column chromatography (

4g

Flash silica gel column, eluent 15-30% tetrahydrofuran/petroleum ether gradient @18 mL/min). The title compound (160mg) was obtained.

MS m/z(ESI):＝630.3[M+H] ⁺

Step 9 Synthesis of (12aS) -10-chloro-8-fluoro-9- (2-fluoro-6-hydroxyphenyl) -7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -3,4,12,12 a-tetrahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepine-2 (1H) -carboxylic acid tert-butyl ester (intermediate 2-11)

Intermediate 2-10(20mg, 31.74. mu. mol) was dissolved in tetrahydrofuran (3mL), borane-tetrahydrofuran solution (1M,634.84uL) was added at 0 ℃ and the reaction was stirred at 40 ℃ for 16 hours. After cooling to 0 ℃, methanol (5mL) was added to the syringe, the mixture was stirred at 25 ℃ for 1 hour, the organic phase was concentrated under reduced pressure (0.01MPa) to remove the solvent, and the residue was purified by preparative thin layer chromatography (silica, tetrahydrofuran: petroleum ether ═ 1: 1). To give the crude title compound (60 mg).

MS m/z(ESI):＝616.1[M+H] ⁺

Step 10 Synthesis of 2- ((S) -10-chloro-8-fluoro-7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -1,2,3,4,12,12 a-hexahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepin-9-yl) -3-fluorophenol (intermediate 2-12)

Intermediate 2-11(52.6mg, 85.38. mu. mol) was dissolved in methylene chloride (4mL), and trifluoroacetic acid (1.54g,13.51mmol) was added to react at 15 ℃ for 1 hour. The solvent was removed by concentration under reduced pressure (0.01 MPa). To give the crude title compound (44.05 mg).

MS m/z(ESI):＝516[M+H] ⁺

Step 11 Synthesis of 1- ((12aS) -10-chloro-8-fluoro-9- (2-fluoro-6-hydroxyphenyl) -7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -3,4,12,12 a-tetrahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepin-2 (1H) -yl) prop-2-en-1-one (Compound 2)

Intermediate 2-12(44mg, 85.27. mu. mol) and triethylamine (69.03mg, 682.20. mu. mol) were dissolved in dichloromethane (2mL), acryloyl chloride (7.72mg, 85.27. mu. mol) was added dropwise at 0 ℃ and the reaction was stirred at 0 ℃ for 1 hour. The solvent was removed by concentration under reduced pressure (0.01 MPa). The residue was purified by preparative high performance liquid chromatography (column: YMC-actual Triart C18100 x 30mm x 5 um; mobile phase: A: water (0.05% ammonia v/v), B: acetonitrile; B%: 45% -65%, 10 min) to give a pair of isomers (atropisomers). Compound 2, isomer 1(1.5mg, LCMS retention time: 1.306min), Compound 2, isomer 2(1.5mg, LCMS retention time: 2.615 min).

LCMS detection conditions: waters Xbridge C1830 × 2.0mm,3.5 um; mobile phase a) 0.04% aqueous solution of trifluoroacetic acid; B) 0.02% trifluoroacetic acid in acetonitrile; gradient elution, increasing mobile phase B from 0% to 95% in 5.8 min; mobile phase B95% was held for 1.1 min; mobile phase B was then reduced to 0% at 6.91 minutes and maintained for 0.09 minutes. The flow rate was 1.0mL/min

MS m/z(ESI):＝570.3[M+H] ⁺

Compound 2 isomer 1 ¹ H NMR(400MHz,METHANOL-d ₄ )δ8.15(dd,J＝1.8,5.0Hz,1H),7.26-7.15(m,1H),7.09(dd,J＝4.6,8.4Hz,1H),6.79(d,J＝13.6Hz,1H),6.68-6.57(m,2H),6.24(d,J＝17.3Hz,1H),5.78(s,1H),4.5-4.43(m,1H),4.41-4.33(m,1H),4.33-4.15(m,1H),4.10-4.00(m,1H),3.91-3.77(m,1H),3.71(m,1H),3.57-3.38(m,1H),3.19-2.96(m,2H),2.88(m,2H),2.58(m,1H),2.23-2.18(s,3H),1.30-1.22(m,3H),1.20-1.10(m,3H)

Compound 2 isomer 2 ¹ H NMR(400MHz,METHANOL-d ₄ )δ8.15(d,J＝5.0Hz,1H),7.26-7.17(m,1H),7.09(d,J＝4.8Hz,1H),6.80(d,J＝8.5Hz,1H),6.70-6.53(m,2H),6.24(d,J＝17.1Hz,1H),5.78(m,1H),4.45-4.35(m,2H),4.33-4.15(m,1H),4.05-3.90(m,1H),3.89-3.78(m,1H),3.75-3.60(m,1H),3.56-3.41(m,1H),3.35-3.13(m,1H),3.12-2.99(m,1H),2.92-2.80(m,2H),2.65-2.59(m 1H),2.25-2.17(m,3H),1.29-1.21(m,3H),1.18-1.05(m,3H)

Example 3, (12aS) -2-acryloyl-10-chloro-8-fluoro-9- (2-fluoro-6-hydroxyphenyl) -7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -1,2,3,4,12,12 a-hexahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepin-6-one (Compound 3)

Step 1 Synthesis of (12aS) -10-chloro-8-fluoro-9- (2-fluoro-6-hydroxyphenyl) -7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -1,2,3,4,12,12 a-hexahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepin-6-one (intermediate 3-1)

To a solution of intermediate 2-10(80mg, 126.97. mu. mol) in dichloromethane (2mL) was added trifluoroacetic acid (14.48mg, 126.97. mu. mol), and the reaction was stirred at 20 ℃ for 2 hours. LCMS showed the starting material reaction was complete. The reaction solution was concentrated to dryness under reduced pressure. To give the crude title compound (81.7 mg).

MS m/z(ESI):＝530[M+H] ⁺

Step 2 Synthesis of (12aS) -2-acryloyl-10-chloro-8-fluoro-9- (2-fluoro-6-hydroxyphenyl) -7- ((2-isopropyl-4-methylpyridin-3-yl) oxy) -1,2,3,4,12,12 a-hexahydro-6H-benzo [ f ] pyrazino [2,1-c ] [1,4] oxazepin-6-one (Compound 3)

Diisopropylethylamine (49.19mg, 380.60. mu. mol) was added to a solution of intermediate 3-1(81.7mg, 126.87. mu. mol) in dichloromethane (3mL) at 0 ℃, the pH was adjusted to 8, a solution of acryloyl chloride (48 mg, 126.87. mu. mol) in dichloromethane was added to the reaction flask with stirring, and the reaction was stirred at 0 ℃ for 20 minutes, then warmed to 20 ℃ and stirred for 1 hour. LCMS showed the starting material reaction was complete. The reaction solution was concentrated to dryness under reduced pressure. The residue was separated and purified by preparative high performance liquid chromatography (basic conditions, column: Boston Prime C18150 × 25mm × 5 um; mobile phase: A: water (0.05% ammonia water v/v), B: acetonitrile; B%: 35% -57%, 7 minutes), and then further purified by supercritical fluid chromatography (conditions: column: DAICEL CHIRALCEL OJ (250mm × 30mm,10 um); mobile phase: B: 0.1% ammonia water in ethanol; B%: 25% -25%) to give a pair of isomers (atropisomers). Compound 3, isomer 1(5.4mg, LCMS retention time: 2.420min), Compound 3, isomer 2(6.2mg, LCMS retention time: 2.699 min).

LCMS detection conditions: waters Xbridge C1830 × 2.0mm,3.5 um; the mobile phase A is 0.04 percent of aqueous solution of trifluoroacetic acid; b, 0.02 percent of acetonitrile solution of trifluoroacetic acid; gradient elution, increasing mobile phase B from 0% to 95% in 5.8 min; mobile phase B95% was held for 1.1 min; mobile phase B was then reduced to 0% at 6.91 minutes and maintained for 0.09 minutes. The flow rate was 1.0mL/min

MS m/z(ESI):＝585[M+H] ⁺

Compound 3 isomer 1 ¹ H NMR(400MHz,Methanol-d ₄ )δ8.15(s,1H),7.25(d,J＝7.9Hz,1H),7.07(s,1H),6.85-6.58(m,3H),6.32(d,J＝16.6Hz,1H),5.84(d,J＝10.5Hz,1H),4.25(s,3H),4.05(s,2H),3.99-3.83(m,1H),3.83-3.71(m,1H),3.69-3.45(m,2H),2.34(s,1.5H),2.16-2.03(m,1.5H),1.37-1.26(m,3H),1.20-1.13(m,3H)

Compound 3 isomer 2 ¹ H NMR(400MHz,Methanol-d ₄ )δ8.16(d,J＝4.9Hz,1H),7.26(s,1H),7.11(s,1H),6.86-6.59(m,3H),6.32(d,J＝16.8Hz,1H),5.84(d,J＝11.4Hz,1H),4.26(s,3H),4.15-4.02(m,2H),4.00-3.83(m,1H),3.81-3.61(m,2H),3.61-3.45(m,1H),2.66(s,1H),2.34(s,2H),2.16-2.00(m,2H),1.42-1.28(m,3H),1.19-1.12(m,3H)

Test example, human non-small cell Lung cancer H358 cell proliferation inhibition test

The brief introduction of the test principle:

KRAS mutations are the drivers of abnormal proliferation of a variety of tumor cells, with KRAS-G12C mutations occurring more highly in non-small cell lung and pancreatic cancers. In vitro, the KRAS-G12C inhibitor is detected to inhibit the cell proliferation by taking human non-small cell lung cancer H358 carrying KRAS-G12C mutation as a cell model. Reference to Experimental methods Janes MR et al, Targeting KRAS Mutant cancer with a covalence G12C-Specific Inhibitor, Cell 2018Jan 25; 172(3):578-589.

Experimental materials and instruments:

NCI-H358 cells were purchased from ATCC (USA);

384 well plates were purchased from corning (usa);

RPMI-1640 medium was purchased from Gibco (USA);

DMEM medium was purchased from gibco (usa);

FBS is available from Gibco (USA);

horse serum (horse serum) was purchased from gibco (usa);

penicilin-streptomycin was purchased from Invitrogen (USA);

1640 complete medium RPMI1640 medium + 10% FBS + 1% Penicillin-streptomycin;

DMEM complete medium: DMEM + 10% FBS + 2.5% horse serum + 1% Penicilin-streptomycin;

celltiter Glo assay kit (2D-CTG reagent) was purchased from Promega (USA);

Echo 550Liquid Handler(Labcyte,USA)；

plate reading apparatus Envision (Perkinelmer, USA).

The experimental method comprises the following steps:

h358 cells were harvested on the first day, counted, cell density adjusted, cell suspension seeded at 40. mu.L into 384 well plates (containing 800 cells/well) placed at 37 ℃ with 5% CO ₂ The culture was carried out overnight in an incubator. The following day, compound dilution and cell treatment. Compound stock solution is diluted to 200 × final concentration in DMSO (e.g., compound final concentration is 1 μ M,10 mM is first diluted to 200 μ M, 2 μ L is taken to 98 μ L DMSO, and 40 μ L200 μ M is taken to source plate after mixing), and compound is added to the first day seeded cell culture plate (200 nL/well, each concentration of test compound) in a gradient using the ECHO550 gradient dilution procedure. The cell culture plate was placed at 37 ℃ in 5% CO ₂ The culture was continued in the incubator for 3 days. On the fifth day, the cell culture plate was removed and incubated with 20. mu.L/well of 2D-CTG reagent at room temperature for 20 min. Put into Envision to read Luminescence signal.

And (3) data analysis:

cell growth inhibition Ratio [% ], [% of Average _ DMSO-Sample ] ], [% of Average _ DMSO-Ratio _ Positive Control ] ] x 100%. 100% inhibition was defined as 1 μ M positive compound (2- ((S) -1-acryloyl-4- (7- (3-hydroxynaphthalen-1-yl) -2- (((S) -1-methylpyrrolidin-2-yl) methoxy) -5,6,7, 8-tetrahydropyrido [3, 4-d)]Pyrimidin-4-yl) piperazin-2-yl) acetonitrile) in the presence of a pharmaceutically acceptable carrier. And (5) performing data analysis processing by using XLFit. The concentration-effect curve was fitted using a non-linear four-parameter curve and the IC of the compound was calculated ₅₀ :

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)*HillSlope))

X:Log of compound concentration

Y:Percent inhibition(％inh)

And (3) test results:

under the test conditions, the tested compound has obvious inhibitory activity on the proliferation of the human non-small cell lung cancer H358 cell with KRAS G12C mutation. The binding activity of the test compounds to KRAS G12C protein is detailed in table 1.

TABLE 1

Compound (I)	IC 50
		Compound 1 isomer 1	271.7nM
Compound 2 isomer 1	396.3nM
		Compound 3 isomer 1	803.3nM

Claims (20)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof:

wherein,

R ¹ selected from the group consisting of ^1a Substituted of the following groups: c ₆ -C ₁₀ Aryl, 5-10 membered heteroaryl;

R ² 、R ³ independently selected from H, F, Cl, Br, I, CN, OH or optionally substituted with R ^2a Substituted of the following groups: NH (NH) ₂ 、C ₁ -C ₆ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7 membered heterocyclyl, C ₁ -C ₆ Alkoxy radical, C ₃ -C ₆ Cycloalkyloxy, 4-7 membered heterocyclyloxy, C ₂ -C ₆ Alkenyl radical, C ₂ -C ₆ An alkynyl group;

R ^4a 、R ^4b selected from H, or R ^4a 、R ^4b Taken together to form ═ O;

R ⁵ is selected from

Q is selected from C (═ O), C (═ NR) ⁸ )、NR ⁹ C(＝O)、S(＝O) ₂ Or NR ⁹ S(＝O) ₂ ；

R ⁸ 、R ⁹ Independently selected from H, C ₁ -C ₆ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7 membered heterocyclyl, C ₆ -C ₁₀ Aryl or 5-10 membered heteroaryl;

when R is ⁵ Is selected from

When R is ⁶ 、R ^7A 、R ^7B Each independently selected from H, F, Cl, Br, I, CN, carboxyl or optionally substituted by R ^6a Substituted of the following groups: c ₁ -C ₆ Alkyl radical, C ₃ -C ₁₀ Cycloalkyl, 4-10 membered heterocyclyl, or R ⁶ 、R ^7A Together with the carbon atoms to which they are attached form partially saturated C ₅ -C ₁₀ Cycloalkyl or 5-10 membered heterocyclyl, said partially saturated C ₅ -C ₁₀ Cycloalkyl or 5-10 membered heterocyclyl is optionally substituted by R ^6a Substitution;

when R is ⁵ Is selected from

When R is ^7C Selected from H or optionally substituted by R ^6a Substituted of the following groups: c ₁ -C ₆ Alkyl radical, C ₃ -C ₁₀ Cycloalkyl, 4-10 membered heterocyclyl;

R ¹⁰ is selected from C ₁ -C ₃ An alkyl group;

n is selected from 0, 1,2 or 3;

l is selected from O, NH and CH ₂ 、C(＝O)、S(＝O) ₂ Or S (═ O);

R ¹¹ selected from the group consisting of ^11a Substituted of the following groups: c ₆ -C ₁₀ Aryl, 5-10 membered heteroaryl;

each R ^1a 、R ^2a 、R ^6a 、R ^11a Independently selected from F, Cl, Br, I, OH, CN, NH ₂ O or optionally R ^b Substituted of the following groups: c ₁ -C ₆ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-10 membered heterocyclyl, C ₁ -C ₆ Alkoxy radical, C ₃ -C ₆ Cycloalkyloxy, 4-10 membered heterocyclyloxy, NH (C) ₁ -C ₆ Alkyl), S (O) ₂ (C ₁ -C ₆ Alkyl groups);

each R ^b Independently selected from F, Cl, Br, I, OH, CN, ═ O, NH ₂ 、SH、C ₁ -C ₆ Alkyl radical, C ₃ -C ₆ Cycloalkyl or 4-7 membered heterocyclyl.

2. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ¹ Selected from the group consisting of ^1a Substituted of the following groups: phenyl, 5-10 membered heteroaryl; preferably, R ¹ Selected from the group consisting of ^1a Substituted of the following groups: phenyl, pyridyl, benzothiazolyl.

3. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ^1a Selected from F, Cl, Br, I, OH, CN, NH ₂ O or optionally R ^b Substituted of the following groups: c ₁ -C ₃ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7 membered heterocyclyl, C ₁ -C ₃ Alkoxy radical, C ₃ -C ₆ Cycloalkyloxy, 4-7 membered heterocyclyloxy, NH (C) ₁ -C ₃ Alkyl groups).

4. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ¹ Is selected from

5. A compound of formula (I) according to claim 1A compound or a pharmaceutically acceptable salt thereof, wherein R is ² 、R ³ Independently selected from H, F, Cl, Br, I, CN, OH or optionally substituted with R ^2a Substituted of the following groups: NH (NH) ₂ 、C ₁ -C ₃ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7 membered heterocyclyl, C ₁ -C ₃ Alkoxy radical, C ₂ -C ₃ Alkenyl radical, C ₂ -C ₃ Alkynyl.

6. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ^2a Selected from halogen, CN, OH, ═ O or C ₁ -C ₃ An alkyl group.

7. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ² Selected from H, halogen, ethynyl, 1-propynyl, cyclopropyl, trifluoromethoxy or difluoromethoxy.

8. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ³ Selected from H, halogen, C ₁ -C ₃ Alkyl or C ₃ -C ₆ A cycloalkyl group.

9. A compound of formula (I), or a pharmaceutically acceptable salt thereof, as claimed in claim 1 wherein Q is selected from C (═ O) or S (═ O) ₂ 。

10. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ⁶ 、R ^7A 、R ^7B 、R ^7C Each independently selected from H, F, Cl, Br, I, CN, C ₁ -C ₃ An alkyl group.

11. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ⁵ Is selected from

12. The compound of formula (I), or a pharmaceutically acceptable salt thereof, as claimed in claim 1, wherein L is selected from O or NH.

13. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ¹¹ Selected from the group consisting of ^11a Substituted of the following groups: phenyl, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl.

14. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ^11a Selected from F, Cl, Br, I, OH, CN, NH ₂ O or optionally R ^b Substituted of the following groups: c ₁ -C ₆ Alkyl radical, C ₃ -C ₆ Cycloalkyl, 4-7 membered heterocyclyl, C ₁ -C ₆ An alkoxy group; preferably, R ^11a Is selected from C ₁ -C ₃ An alkyl group; more preferably, R ^11a Selected from methyl or isopropyl.

15. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is ¹¹ Is selected from

16. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

17. the compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

18. the compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

19. a pharmaceutical composition comprising a compound of any one of claims 1 to 18, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable adjuvant.

20. Use of a compound of any one of claims 1 to 18, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 19, in the manufacture of a medicament for preventing or treating a disease associated with KRAS G12C.